We applied urease-induced silver metallization on the surface of gold nanorods (AuNR) to improve colorimetric ELISA for the rapid and sensitive detection of Salmonella enterica Choleraesuis. To this end, we introduced a biotin-streptavidin system as a bridge to determine the correlation between urease and S. enterica Choleraesuis concentrations. The captured urease can catalyze the hydrolysis of urea into carbon dioxide and ammonia, and the generated ammonia can then induce the deposition of silver shell on the surface of AuNR in the presence of silver nitrate and glucose. With the decreased aspect ratio (length divided by width) of AuNR, a multicolor change of AuNR solution and a significant blue shift in the longitudinal localized surface plasmon resonance absorption peak (Δλmax) of AuNR were obtained at the target concentrations of 1.21 × 101 to 1.21 × 108 cfu/mL. Consequently, the detection limits of our proposed colorimetric ELISA were as low as 1.21 × 102 cfu/mL for qualitative detection with naked eyes, and 1.21 × 101 cfu/mL for quantitative detection, in which changes in Δλmax of AuNR were recorded with a microplate reader. These values were at least 2 to 3 orders of magnitude lower than those obtained with conventional horseradish peroxidase-based ELISA. The analytical performance of our developed colorimetric ELISA in terms of selectivity, accuracy, reliability, and practicability were investigated by analyzing S. enterica Choleraesuis-spiked pasteurized whole milk samples.
Keywords: ELISA; Salmonella enterica Choleraesuis; colorimetric detection; gold nanorod; urease metallization.
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